Difference between revisions of "Part:BBa K5124011"
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===Characterisation=== | ===Characterisation=== | ||
− | This crRNA sequence was taken from the paper by Moreno-Mateos <i>et al.</i> [2]. It was synthesised by IDT as part of one of five composite parts (BBa_K5124035 to BBa_K5124040) each containing: a 3’ spacer sequence (BBa_K5124018 to BBa_K5124023), 5’ T7 promoter | + | This crRNA sequence was taken from the paper by Moreno-Mateos <i>et al.</i> [2]. It was synthesised by IDT as part of one of five composite parts (BBa_K5124035 to BBa_K5124040) each containing: a 3’ spacer sequence (BBa_K5124018 to BBa_K5124023), 5’ T7 promoter [https://parts.igem.org/Part:BBa_I719005 BBa_I719005] and BioBrick compatible prefix and suffixes. The g-block was cloned into a high copy plasmid (origin of replication from pUC18 [3]) carrying an ampicillin selection marker. |
Please see composite parts: | Please see composite parts: | ||
− | + | <br>[https://parts.igem.org/Part:BBa_K5124035 K5124035]</br> | |
− | + | <br>[https://parts.igem.org/Part:BBa_K5124036 K5124036]</br> | |
− | + | <br>[https://parts.igem.org/Part:BBa_K5124037 K5124037]</br> | |
− | + | <br>[https://parts.igem.org/Part:BBa_K5124038 K5124038]</br> | |
− | + | <br>[https://parts.igem.org/Part:BBa_K5124039 K5124039]</br> | |
− | + | <br>[https://parts.igem.org/Part:BBa_K5124040 K5124040]</br> | |
for further usage and results. | for further usage and results. | ||
Revision as of 13:09, 22 August 2024
Cas12a crRNA
Usage and Biology
The Exeter iGEM 2024 team are designing a rapid detection system for Bovine Tuberculosis (bTB) using CRISPR-Cas detection systems.
This basic part codes for the CRISPR-RNA (crRNA) repeat sequence found in the class II, type V CRISPR loci of Lachnospiraceae bacterium ND2006 [1]. This sequence is combined with one of five spacer sequences that are complimentary to our target bovine TB DNA. Once transcribed into RNA, the 23-nucleotide repeat sequence folds into a single hairpin loop, which is recognised and bound by LbCas12a, leaving the 20-nucleotide spacer sequence free to bind to the target DNA (Figure 1).
Characterisation
This crRNA sequence was taken from the paper by Moreno-Mateos et al. [2]. It was synthesised by IDT as part of one of five composite parts (BBa_K5124035 to BBa_K5124040) each containing: a 3’ spacer sequence (BBa_K5124018 to BBa_K5124023), 5’ T7 promoter BBa_I719005 and BioBrick compatible prefix and suffixes. The g-block was cloned into a high copy plasmid (origin of replication from pUC18 [3]) carrying an ampicillin selection marker.
Please see composite parts:
K5124035</br>
K5124036</br>
K5124037</br>
K5124038</br>
K5124039</br>
K5124040</br>
for further usage and results.
References
[1] Zetsche B, Gootenberg JS, Abudayyeh OO, Slaymaker IM, Makarova KS, Essletzbichler P, et al. Cpf1 is a single RNA-guided endonuclease of a class 2 CRISPR-Cas system. Cell. 2015 Oct 22; 163(3):759-71.
[2] Moreno-Mateos MA, Fernandez JP, Rouet R, Vejnar CE, Lane MA, Mis E, et al. CRISPR-Cpf1 mediates efficient homology-directed repair and temperature-controlled genome editing. Nat Commun. 2024 Dec 8; 8:1-9.
[3] Vieira J, Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct; 19(3):259-68.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]